Remminghorst Uwe, Rehm Bernd H A
Institute of Molecular BioSciences, Massey University, Private Bag 11222, Palmerston North, New Zealand.
FEBS Lett. 2006 Jul 10;580(16):3883-8. doi: 10.1016/j.febslet.2006.05.077. Epub 2006 Jun 16.
Here the putative alginate biosynthesis gene alg44 of Pseudomonas aeruginosa was functionally assigned. Non-polar isogenic alg44 deletion mutants of P. aeruginosa were generated and did neither produce alginate nor released free uronic acids. No evidence for alginate enrichment in the periplasm was obtained. Alginate production was restored by introducing only the gene alg44. PhoA fusion protein analyses suggested that Alg44 is a soluble protein localized in the periplasm. Hexahistidine-tagged Alg44 was detected by immunoblotting. The corresponding 42.6 kDa protein was purified and identified by MALDI/TOF-MS analysis. Alg44 might be directly involved in alginate polymerization presumably by exerting a regulatory function.
在此对铜绿假单胞菌假定的藻酸盐生物合成基因alg44进行了功能鉴定。构建了铜绿假单胞菌的非极性同基因alg44缺失突变体，该突变体既不产生藻酸盐，也不释放游离糖醛酸。未获得周质中藻酸盐富集的证据。仅导入基因alg44可恢复藻酸盐的产生。PhoA融合蛋白分析表明，Alg44是一种定位于周质的可溶性蛋白。通过免疫印迹检测到带六聚组氨酸标签的Alg44。纯化了相应的42.6 kDa蛋白，并通过基质辅助激光解吸电离飞行时间质谱（MALDI/TOF-MS）分析进行鉴定。Alg44可能通过发挥调节功能直接参与藻酸盐聚合。
